Alteration by heat activation of enzymes localized in spore coats of Bacillus cereus.
Heat activation (70 degrees C for 20 min) resulted in alteration in structural proteins and enzymes found in Bacillus cereus spore coats. The three notable changes were increased glycosylation of coat proteins, alteration in polypeptide pattern on sodium dodecyl sulfate - polyacrylamide gels, and an increase in free SH groups of proteins. About three polypeptides leaked out in small quantities from the spore coats during heat activation. The extraction of five spore coat associated enzyme activities was followed during the coat stripping procedures, which left the cortex and core intact. Two of these activities, L-alanine dehydrogenase and purine nucleoside hydrolase, were solubilized when the undercoat was extracted by 1,4-dithioerythritol (DTE) at pH 9.8. Three other activities, a protease, a corticolytic enzyme, and purine nucleoside phosphorylase, were solubilized by both DTE alone and DTE plus urea at pH 9.8. The DTE plus urea extraction removed the two more insoluble coat layers, the outer cross-patch, and the inner pitted layers. Mutants deficient in the cross-patch layer contained normal amounts of the protease, corticolytic, and purine nucleoside phosphorylase activities suggesting their association with the pitted layer. In intact spores all five enzymes were found to be stable to the heat activation treatment. However, extracted and partially purified preparations of protease, purine nucleoside phosphorylase, and L-alanine dehydrogenase were heat sensitive. Similar preparations of corticolytic enzyme and purine nucleoside hydrolase were stable to the heat activation conditions.
D050788:Alanine Dehydrogenase; D000594:Amino Acid Oxidoreductases / Q000378:metabolism; D001409:Bacillus cereus / Q000201:enzymology* / Q000502:physiology; D004789:Enzyme Activation; D006358:Hot Temperature; D009699:N-Glycosyl Hydrolases / Q000378:metabolism; D010447:Peptide Hydrolases / Q000378:metabolism; D011683:Purine-Nucleoside Phosphorylase / Q000378:metabolism; D013171:Spores, Bacterial / Q000201:enzymology
D000594:Amino Acid Oxidoreductases; D050788:Alanine Dehydrogenase; D011683:Purine-Nucleoside Phosphorylase; D009699:N-Glycosyl Hydrolases; C030681:purine nucleosidase; D010447:Peptide Hydrolases